Necroinflammatory Activity Research Articles

Risk of hepatitis B virus reactivation in hepatitis B virus+hepatitis C virus-co-infected patients with compensated liver cirrhosis treated with ombitasvir, paritaprevir/r+dasabuvir+ribavirin.

Hepatitis B virus may reactivate in patients with chronic hepatitis C treated with direct-acting antivirals. The aim of this study was to investigate the risk of hepatitis B virus (HBV) reactivation in HBV+hepatitis C virus (HCV)-co-infected patients with compensated liver cirrhosis treated with paritaprevir/ombitasvir/ritonavir, dasabuvir with ribavirin. We reviewed prospectively gathered data from a national cohort of 2070 hepatitis C virus patients with compensated liver cirrhosis who received reimbursed paritaprevir/ombitasvir/r, dasabuvir with ribavirin for 12weeks from the Romanian National Health Agency during 2015-2016. Twenty-five patients in this cohort were HBs antigen positive (1.2%); 15 untreated with nucleotide analogues agreed to enter the study. These patients were followed up: ALT monthly, serology for HBV and DNA viral load at baseline, EOT and SVR at 12weeks. Hepatitis B virus (HBV)-co-infected patients were all genotype 1b and 52% females, with a median age of 60years (51÷74); 76% were pretreated with peginterferon+ribavirin; 72% were with severe necroinflammatory activity on FibroMax assessment; 40% presented comorbidities; and all were HBe antigen negative. Hepatitis C virus (HCV) SVR response rate was 100%. Hepatitis B virus (HBV)-DNA viral load was undetectable in 7/15 (47%) before therapy, and for the other 8 patients, it varied between below 20 and 867IU/mL. Five patients (33%) presented virological reactivation (>2 log increase in HBV-DNA levels) during therapy. One patient presented with hepatitis associated with HBV reactivation, and two started anti-HBV therapy with entecavir. Hepatitis B virus (HBV) virological reactivation was present in 33% in our patients. Generally, HBV-DNA elevations were mild (<20000IU/mL); however, we report one case of hepatitis associated with HBV reactivation.

Soluble CD163 and mannose receptor associate with chronic hepatitis B activity and fibrosis and decline with treatment.

Liver macrophages are activated in chronic hepatitis B virus (CHB) infection and play a pivotal role in hepatic inflammation and fibrosis. However, their role during antiviral treatment is unclear. The soluble (s) macrophage activation markers, sCD163 and mannose receptor (sMR), are released during liver damage, and their serum levels reflect liver disease severity and portal hypertension. We aimed to investigate associations between sCD163 and sMR and histopathological activity and fibrosis and changes in sCD163, sMR, and hepatic CD163-expression following antiviral treatment in CHB patients. We assessed Ishak histological necroinflammatory activity and fibrosis scores in liver biopsies from 254 CHB patients and serially in 71 patients before and after nucleoside-analogue treatment. Liver CD163-expression was semi-quantitatively determined by immunohistochemistry and serum sCD163 and sMR measured by enzyme-linked immunosorbent assays. Before treatment, the mean levels of sCD163 and sMR were 3.57 (SD 1.72) mg/L and 0.35 (0.12) mg/L. sCD163 and sMR increased with histological inflammatory activity (sCD163: r=0.46, P<0.00001; sMR: r=0.48, P<0.00001) and correlated positively with fibrosis (sCD163: OR 1.16, 95% CI:1.03-1.31; sMR: OR 1.34, 95% CI:1.13-1.59); both were markers of fibrosis independent of other biochemical parameters and risk factors. Antiviral treatment significantly reduced sCD163 (3.76 [1.46] vs 2.31 [0.95], P<0.00001), sMR (0.37 [0.1] vs 0.29 [0.07], P<0.00001) and hepatic CD163-expression (P=0.0002). The macrophage activation markers sCD163 and sMR were associated with activity and fibrosis in liver biopsies from CHB patients. Both serum markers decreased with antiviral treatment, along with decreased hepatic CD163 expression.

Biopsy Proven Fibrosis in Non-Alcoholic Fatty Liver Disease: An Analysis of Risk Factors

Non-alcoholic fatty liver disease (NAFLD) is emerging as an important cause of liver disease in India. NAFLD is characterized by hepatic steatosis in absence of a significant alcohol use or other known liver disease. Non-alcoholic steatohepatitis (NASH) is a progressive form of NAFLD which deserves particular attention because it is more prone for development of fibrosis. Liver biopsy is the gold standard for diagnosis of NASH by evaluating necroinflammatory activity and stages of fibrosis. The aim of the study was to analyze liver biopsy specimens and identify risk factors associated with fibrosis in patients of NAFLD in eastern coastal India. A total of 216 subjects with fatty liver in ultrasonography (USG) were selected for needle biopsy. Those NAFLD cases showing fibrosis in biopsy were analyzed for risk factors association. Definite NASH was diagnosed in 50 (23.14%), borderline NASH in 66 (30.55%) and not NASH in 100 (46.39%) of cases. Those patients with fibrosis (22%) were taken as cases and those without fibrosis (78%) were taken as controls for risk factor analysis. Age>40 [odds ratio (OR) 2.01 (1.09-4.04)], female gender [OR 2.74 (1.24-6.05)], body mass index (BMI)>23 [OR 15.36 (4.59-51.37)] and moderate fatty change in USG [OR 1.89 (1.01-3.62)] were observed as risk factors for progression to fibrosis in NAFLD cases. Older age, females, obesity and moderate fatty liver on USG are risk factors for development of fibrosis in patients with NAFLD. Patients with these risk factors should be selected for liver biopsy and to be kept for close follow-up.

The Clinical Features of Patients with Chronic Hepatitis C Virus Infections Are Associated with Killer Cell Immunoglobulin-Like Receptor Genes and Their Expression on the Surface of Natural Killer Cells.

Killer cell immunoglobulin-like receptor (KIR) genes are known to play a role in the acute phase of hepatitis C virus (HCV) infection. The present study investigated their roles in chronic HCV (CHCV) infection by analyzing the phenotypes and function of natural killer (NK) and T cells that express KIRs. T cells from CHCV patients showed a more differentiated phenotype, and NK cells exhibited an activated profile. These observations are consistent with the increased expression of the degranulation marker CD107a observed after PMA stimulation. We explored the correlations between the expression of KIR genes and lectin type-C receptors with clinical factors that predict progression to fibrosis and cirrhosis. The expression levels of KIR2DS3 and the functional alleles of KIR2DS4-FL were increased in patients with intermediate and high viral loads. Homozygous KIR2DS4 was also associated with the presence of cirrhosis. In the group of individuals with a shorter infection time who developed cirrhosis, we detected decreased expression of KIR3DL1 in CD56dim NK cells in the presence of its ligand. Similarly, in the group of patients with late CHCV infections complicated with cirrhosis, we detected lower expression of the strong inhibitory receptor NKG2A in CD56bright NK cells. We also detected an increase in NKG2C expression in CD56dim NK cells in CHCV patients who displayed high necroinflammatory activity. Decreased KIR3DL2 expression in CD56dim and CD56bright NK cells was associated with a high body mass index, and KIR3DL2 expression may be one factor associated with the more rapid progression of CHCV to fibrosis in patients.

Impact of Different Sources of Infection on Therapy Response in Chronic Hepatitis C.

Introduction:Prior to the 1990s, the most common sources of HCV infections were blood transfusions, unsafe injections and I.V drug use. Screening of blood products for HCV has eradicated transfusion-transmitted hepatitis C in most countries since 1992–in Bosnia and Herzegovina, however, since 1995, due to the war.Aim:To investigate the impact of the source of HCV infection on the therapeutic response in patients treated for chronic HCV infection with dual combined therapy.Methods:We diagnosed chronic HCV infections amongst 246 patients over a period of five years and selected them according to the reported source of infection. Pegylated interferon alfa 2a or alfa 2b with ribavirin was administered during the time that was genotype-dependent. HCV RNA levels in sera were measured by real time PCR. Liver histology was evaluated in accordance with the level of necroinflammation activity and the stadium of fibrosis.Results:Regardless of the genotype of the virus and the source of infection, SVR was achieved in 67% of the patients. Therapeutic response (ETR) was not achieved in 25% of the patients who were infected with an untested blood transfusion and 6% of the patients who had had wartime surgery. Amongst the different sources of infections, patients with a war-surgery source of infection responded better to therapy than those with a blood transfusion source of infection (p = 0.023). A blood transfusion source of infection implies a larger fibrosis stage than in blood donors; (g = 1.177; s2 = 0.577). A blood transfusion source of infection implies a significantly larger necroinflammatory activity than in blood donors; (g = 1.456; s2 = 0.618).Conclusions:An untested blood transfusion was a significant risk factor for more advanced liver diseases in regards to necroinflammatory activity and the fibrosis stage. This source of infection was also a risk factor for low responses to antiviral therapy. At the same time, I.V. drug users had more progressive necroinflammatory activity, but a high therapeutic response to antiviral therapy.

Clinical and Virologic Characteristics of HIV-1 Positive Patients with Delta Hepatitis

Background and Aim: Hepatitis Delta Virus (HDV) infection has been mainly studied in HIV negative patients, while data on HIV-1 positive patients are limited. We investigated the virological pattern as well as biochemical and clinical features of liver disease and immune status in HIV-1 positive patients with delta hepatitis. Their clinical characteristics were compared with those of anti-HDV negative, hepatitis B surface antigen (HBsAg) positive/HIV+ patients. Methods: This retrospective study included HBsAg positive subjects with anti-HDV serology available, during the period 2010-2017. Biochemical and virological parameters were obtained at last visit in 2017 for each patient. Potential determinants for HDV positivity were examined by applying multivariate regression model. Results: Of 78 HBsAg positive patients 19 (24.4%) were found anti-HDV+. Anti-HDV+ patients were more frequently intra venous drug users, anti-HCV positive and HBV e antigen (HBeAg) negative. Additionally, the patients had more severe liver disease and necro inflammatory activity (assessed by transient elastography and transaminases levels, repectively) than the counterpart of anti-HDV- patients. A suppressive effect of HDV over HCV was also revealed in anti-HDV+ subjects. By multivariate analysis, years of ART (OR 1.22; CI 0.986-1.43, p=0.014) and sexual exposure vs. IVDU (OR 0.08; CI 0.556-0.986, p=0.004) were independently associated with anti-HDV positivity. Conclusion: Our data underlines the need for continuing prevention program that includes HBV vaccination, screening and monitoring in population at high risk, as well as development of an alternative treatment option for HDV.

Abstract B78: Serum microRNA panel to diagnose important liver diseases

Abstract MicroRNA-21 (miR-21) has been described as upregulated in tissue and serum samples from patients with tumor diseases such as hepatocellular carcinoma. In addition, circulating miR-21 has been reported as a marker of necroinflammatory activity in patients with hepatitis C virus (HCV) and HCV-induced hepatocellular carcinoma (HCC). Thus, the expression of miR-21 by reverse transcription quantitative real-time PCR (RT-qPCR) in serum samples (n = 216) from patients with hepatic cirrhosis (n = 187) compared with control group (n = 29) was evaluated. Initially, peripheral blood was collected in tubes containing separating gel and clot activator and centrifuged at 820 × g for 10 min at room temperature. Then, the supernatants were transferred to new microtubes and the samples were centrifuged at 16,000 × g for 10 min to precipitate cell debris. Next, total serum RNA was isolated from 500 μL of serum from each sample and eluted in 50 μL of RNase-free water using a mirVanaTM PARISTM kit (AM1556, Ambion, Carlsbad, CA), according to the manufacturer's instructions. After that, total RNA quality obtained was evaluated by NanoVueTM Plus spectrophotometer (GE Healthcare, Buckinghamshire, UK) and the RT-qPCR was done using Taqman microRNA assays (Applied Biosystems, Foster City, CA). In the present work, the expression of miR-16 was not differentially expressed between healthy (Cq = 24.7 ± 3.0) and cirrhosis groups (Cq = 25.3 ± 3.0; P = 0.2741). Therefore, miR-16 was used as internal control to evaluate the expression of miR-21. The relative expression of miR-21 found was not statistically significant considering the control (1.0 ± 0.6) and cirrhosis groups (2.0 ± 3.5; P = 0.1478). The low expression of miR-21 indicates the patients did not yet develop a malignant disease or necroinflammatory activity. To better evaluate hepatic function, miR-34a, miR-122, and miR-885-5p were also analyzed, because these miRNAs have already been described as potential biomarkers for detection and assessment of liver diseases. The relative expressions found were significantly different for miR-34a (1.0 ± 1.5 vs 3.5 ± 8.1, P = 0.0077), miR-122 (1.0 ± 10.1 vs 1.2 ± 33.6, P &amp;lt; 0.0001) and miR-885-5p (1.0 ± 4.8 vs 1.9 ± 12.8, P = 0.0001). These miRNAs were found to be upregulated with fold-change for miR-34a, miR-122, and miR-885-5p of 4.1, 7.1, and 3.2, respectively. Due to the fact that these miRNAs were previously studied separately in independent studies, we applied a logistic regression model to obtain a miRNA panel not yet described. The logit model, logit(p = Cirrhosis) = 0.59 + 1.57 x miR-34a + 0.50 x miR-122 - 0.14 x miR-885-5p, was used to construct the ROC curve and, thus, to evaluate the diagnostic accuracy for the established miRNA panel. The area under the ROC curve (AUC) for the miRNA panel was 0.859 (95% confidence interval, 0.792-0.912; sensitivity: 84.8%; specificity: 73.7%; P &amp;lt; 0.0001), providing the high diagnostic accuracy for cirrhosis. A serum microRNA panel having considerable clinical value in cirrhosis diagnosis, chronic hepatitis viral, and hepatocellular carcinoma was found, thus allowing patients not to lose the window of treatment being benefited with a more appropriate therapy. Citation Format: Alex Evangelista Amaral, Júlia Cisilotto, Michele Patrícia Rode, Evelyn Winter, Adny Henrique Silva, Jelver Sierra-Restrepo, Leonardo de Lucca Schiavon, Tânia Beatriz Creczynski-Pasa. Serum microRNA panel to diagnose important liver diseases [abstract]. In: Proceedings of the AACR International Conference held in cooperation with the Latin American Cooperative Oncology Group (LACOG) on Translational Cancer Medicine; May 4-6, 2017; São Paulo, Brazil. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(1_Suppl):Abstract nr B78.

Centrilobular ductular reaction correlates with fibrosis stage and fibrosis progression in non-alcoholic steatohepatitis

There is increasing interest in the role of ductular reaction as part of the pathogenesis and characteristic histology of non-alcoholic steatohepatitis. However, earlier studies did not separately assess the contribution of periportal and centrilobular zone ductular reaction over the spectrum of non-alcoholic steatohepatitis, and their clinical significance remains unclear. We herein analyzed the character of ductular reaction in each hepatic zone in non-alcoholic steatohepatitis biopsies and for the first time evaluated the prognostic value of ductular reaction in baseline biopsies as a predictor of progression of fibrosis in subsequent biopsies. A total of 90 non-alcoholic steatohepatitis liver biopsies were included in the cohort. The relationships among ductular reaction, grade, stage, and other common histopathologic findings in non-alcoholic steatohepatitis were analyzed in a cross-sectional manner. Among these patients, a total of 47 patients underwent sequential liver biopsies in the absence of effective treatment. The frequency of ductular reaction and the other histopathologic parameters in the initial biopsies were analyzed as predictors of progression of fibrosis in the second biopsies in a longitudinal analysis. Centrilobular ductular reaction was identified in 90% of patients and 38% of centrilobular zones. The prevalence of centrilobular ductular reaction increased as non-alcoholic steatohepatitis grade increased (P=0.0002) and also as stage of fibrosis increased (P<0.0001) in the cross-sectional study. In the longitudinal study, the frequency of centrilobular ductular reaction in the initial biopsies was significantly higher in the group of progressors and correlated with the rate of fibrosis progression (P=0.02). Centrilobular ductular reaction is common in non-alcoholic steatohepatitis and its presence correlates significantly with increasing necroinflammatory activity and fibrosis stage. Development of centrilobular ductular reaction appears to predict progression of fibrosis in subsequent biopsies.

Liver HLA-E Expression Is Associated with Severity of Liver Disease in Chronic Hepatitis C

Hepatitis C virus (HCV) can escape from innate and adaptive immunity, making the immune response ineffective. Human leukocyte antigen E (HLA-E) might regulate the antiviral function of immune response and contribute to the persistence of HCV and the severity of liver disease. This study aimed to evaluate the expression of HLA-E in the liver and its association with the severity of liver disease in HCV patients. We performed a retrospective analysis of liver biopsies from 125 HCV patients and from 20 control subjects without liver disease. Liver biopsies were reviewed and classified according to severity of fibrosis and inflammatory activity. The pathologist assessed the magnitude of HLA-E expression in a semiquantitative way, attributing scores from 0 to 3. Immunohistochemistry showed positive for HLA-E in hepatocyte and Kupffer cells. The rate of HLA-E positivity in hepatocytes and Kupffer cells was significantly higher in HCV patients compared to controls. The liver samples classified as severe fibrosis and necroinflammatory activity presented greater expression of HLA-E on Kupffer cells and hepatocytes, with a significant linear association. It indicates that HLA-E expression may have an immunomodulatory effect and a possible role in the severity of liver disease in chronic hepatitis C.

Relationship between hepatic progenitor cells and stellate cells in chronic hepatitis C genotype 4.

Hepatitis C virus (HCV) infection represents a major health problem in many areas of the world, especially Egypt. Hepatic progenitor cells (HPCs) and hepatic stellate cells (HSCs) have been implicated in fibrosis progression in chronic HCV. The aim of this study was to investigate the role of HPCs and HSCs in chronic HCV infection and the relationship between both cell types. This retrospective study was conducted on 100 chronic HCV patients. Immunohistochemistry was performed on liver tissue sections for cytokeratin 19 (progenitor cell markers), smooth muscle actin (stellate cell markers), matrix metalloproteinase-9 (MMP-9), and transforming growth factor beta (TGF-ß). The necroinflammatory activity was significantly related to the number of isolated HPCs and TGF-ß expression (p = 0.003 and p = 0.001 respectively). Advanced stages of fibrosis showed significantly increase number of HPCs (p = 0.001), higher ratio of HSCs (p = 0.004), more expression of TGF-ß (p = 0.001) and MMP-9 (p = 0.001). There was a significant direct correlation between immunoexpression of HPCs and HSCs for isolated cells (r = 0.569, p = 0.001) and ductular reaction (r = 0.519, p = 0.001). Hepatic progenitor cells and stellate cells play a significant role in the development and progression of fibrosis in chronic HCV. More interestingly, the significant direct correlation between HPCs and HSCs suggests a synergistic interrelation.

MR elastography is effective for the non-invasive evaluation of fibrosis and necroinflammatory activity in patients with nonalcoholic fatty liver disease

ObjectivesTo evaluate the performance of magnetic resonance elastography (MRE) in diagnosing and staging hepatic fibrosis in patients with histologically confirmed nonalcoholic fatty liver disease (NAFLD) and in distinguishing simple steatosis from nonalcoholic steatohepatitis (NASH). MethodsNinety subjects (49 NAFLD patients and 41 healthy volunteers) were prospectively enrolled. Liver stiffness measured by MRE was correlated with the grade of fibrosis and/or inflammation determined by liver biopsy. Correlations, ROC (receiver operator characteristic) curves and diagnostic performance were evaluated. The study was approved by the local ethics committee. ResultsThe area under the ROC curve (AUROC) of MRE in discriminating healthy from NAFLD individuals was 0.964 (P<0.0001), and that for distinguishing advanced (F3-F4) from absent/mild fibrosis (F0-F2) was 0.928 (P<0.0001). The use of a threshold >4.39 kPa resulted in a sensitivity of 90.9% and a specificity of 97.3% for diagnosing advanced fibrosis. For discriminating NASH from simple steatosis, the AUROC was 0.783 (P<0.0001), and the threshold, 3.22 kPa. ConclusionsMRE is an effective, non-invasive method for detecting/staging hepatic fibrosis in NAFLD. This method has good performance in discriminating normal from NAFLD subjects and between the extreme grades of fibrosis. NAFLD patients with inflammation and without fibrosis have higher liver stiffness than those with simple steatosis.

Improvement of liver stiffness in patients with hepatitis C virus infection who received direct‐acting antiviral therapy and achieved sustained virological response

There is insufficient research on whether direct-acting antiviral (DAA) therapy can improve liver fibrosis in patients with chronic hepatitis C virus (HCV). We evaluated sequential changes in liver stiffness using shear wave elastography in patients with HCV who received DAA therapy. A total of 210 patients with HCV who received daclatasvir and asunaprevir therapy and achieved sustained virological response (SVR) were analyzed. Liver stiffness, as evaluated by shear wave elastography, and laboratory data were assessed before treatment (baseline), at end of treatment (EOT), and at 24weeks after EOT (SVR24). Alanine aminotransferase levels (ALT) decreased over time, and there were significant differences between baseline and EOT and between EOT and SVR24. Although platelet counts did not significantly differ between baseline and EOT, they increased significantly from EOT to SVR24. The median (interquartile range) liver stiffness values at baseline, EOT, and SVR24 were 10.2 (7.7-14.7), 8.8 (7.1-12.1), and 7.6 (6.3-10.3) kPa, respectively (P<0.001, baseline vs EOT; P<0.001, EOT vs SVR24). Additionally, in patients with ALT ≤30 (indicating low necroinflammatory activity in the liver) and Fibrosis-4 index >2.0 (n=75), the liver stiffness values at baseline, EOT, and SVR24 were 9.6 (7.7-15.2), 9.2 (7.3-12.1), and 7.7 (6.3-10.1) kPa, respectively (P<0.001, baseline vs EOT; P<0.001, EOT vs SVR24). These results suggest that early improvement of liver stiffness starts during the administration of DAAs in patients who achieve SVR, and this effect is particularly pronounced in patients with progressive liver fibrosis.

ADAR1 polymorphisms are related to severity of liver fibrosis in HIV/HCV-coinfected patients

The adenosine deaminase acting on RNA (ADAR1) gene is an interferon-stimulated gene involved in liver injury protection. Our aim was to analyze the association of polymorphisms within this gene with the severity of liver disease in European HIV/HCV-coinfected patients. We performed a cross-sectional study in 220 patients that underwent a liver biopsy. Five SNPs in the ADAR1 gene (rs1127326, rs1127317, rs1127314, rs1127313, rs2229857) were genotyped by GoldenGate assay. The outcome variables were fibrosis stage and necroinflammatory activity grade by METAVIR-score, aspartate aminotransferase to platelet ratio index (APRI), FIB-4 index, and fibrosis progression rate (FPR). In multivariate analysis, fibrosis progression rate (FPR) (aAMRs = 0.97) decreased in a dose-dependent manner with the presence of rs2229857_T, rs1127313_G, rs1127314_G and rs1127317_G; while rs1127326_T allele had only significant associations with FIB-4 (aAMRs ≤ 0.63) and FPR (aAMRs ≤ 0.97). Moreover, carriers of rs2229857_T, rs1127314_G, rs1127317_G, and rs1127326_T alleles were protected against advanced fibrosis (F ≥ 3) (adjusted ORs (aORs) ≤ 0.44), APRI ≥ 1.5 (aORs ≤ 0.33), and FPR ≥ 0.075 (aORs ≤ 0.45). rs1127313_G carriers showed lower odds of having F ≥ 3 (aORs = 0.39), FIB4 ≥ 3.25 (aOR = 0.22) and FPR ≥ 0.075 (aORs = 0.44). In conclusion, ADAR1 polymorphisms protected against severe liver disease in HIV/HCV-coinfected patients. These results could be used to improve therapeutic decision-making in clinical practice.

Prospective Comparison of the Diagnostic Performance of Magnetic Resonance Elastography with Acoustic Radiation Force Impulse Elastography for Pre-operative Staging of Hepatic Fibrosis in Patients with Hepatocellular Carcinoma

The purpose of this study was to compare the diagnostic accuracy of magnetic resonance (MR) elastography with that of acoustic radiation force impulse (ARFI) elastography for pre-operative staging of hepatic fibrosis in patients with hepatocellular carcinoma. We prospectively enrolled 77 patients who were scheduled to undergo hepatectomy for hepatocellular carcinoma. Pre-operative MRE and ARFI elastography examinations were performed on the same day, and liver stiffness/velocity values were determined. Fibrosis stage and necro-inflammatory activity of resected specimens were determined histopathologically using the METAVIR scoring system. Correlations between MRE and ARFI elastography findings and histologic findings were determined by receiver operating characteristic (ROC) analysis. Correlation of MRE was excellent and correlation of ARFI elastography was good with fibrosis stage. MRE had better diagnostic performance than ARFI elastography in estimating substantial fibrosis (F2), severe fibrosis (F3) and cirrhosis (F4). The optimal cutoff value and the area under the ROC curve (AUROC) were determined using ROC curve analysis. The highest Youden index was used as a criterion for selecting the optimal cutoff value. ROC analysis revealed that MRE discriminated advanced stages of fibrosis (F ≥ 2) well in patients with hepatocellular carcinoma at a cutoff value of 3.0 kPa with an AUROC value of 0.93, and ARFI elastography did so at a cutoff value of 1.77 m/s with an AUROC value of 0.81 for predicting advanced stages of fibrosis (F ≥ 2). In conclusion, MRE is a more accurate imaging modality than ARFI elastography in estimating advanced stages of fibrosis and cirrhosis.

Evaluation of CD44 and CD133 as markers of liver cancer stem cells in Egyptian patients with HCV-induced chronic liver diseases versus hepatocellular carcinoma.

BackgroundCancer stem cells (CSCs) play a critical role in tumor development, progression, metastasis and recurrence.AimTo evaluate hepatic expression of CD44 and CD133 in Egyptian patients with HCV-induced chronic liver diseases and hepatocellular carcinomas (HCCs), and to assess its correlation with inflammatory activity scores, stages of fibrosis (in chronic hepatitis with or without cirrhosis) and grades of HCC.MethodsThis prospective case-control study was conducted on eighty subjects who attended the Tropical Diseases Department, Al-Azhar University Hospital, and in collaboration with Theodor Bilharz Research Institute (2014–2016). They were divided as follows: A) Control healthy group: Ten individuals with serologically negative HCV-Ab and HBsAg, and histopathologically normal liver, B) Seventy patients subdivided into 3 groups; Twenty subjects each, as: HCV-Ab+ non-cirrhotic, HCV-Ab+ cirrhotic and HCC. Necroinflammatory activity and fibrosis in non-neoplastic liver biopsies were scored according to the METAVIR scoring system. CD44 and CD133 immunostaining was evaluated in all groups semi-quantitatively using H score. Statistical analysis was performed by SPSS version 22, using independent-samples t-test.ResultsOur study showed a significant increase of mean CD44 & CD133 expression values with disease progression among the groups (p<0.05). Their expressions increased significantly with the inflammatory activity scores and stages of fibrosis, reaching the highest values in A3F4 score compared to A1F1 (p<0.05). Moreover, there was a significant increase of their expressions across HCC grades (p<0.05), however with no significant correlation with focal lesions size.ConclusionCSCs clusters exhibiting CD133+ and/or CD44+ profiles were identified in chronic hepatitis, liver cirrhosis and HCC. CD133 and CD44 expressions significantly corresponded to the increased inflammatory activity, fibrosis stages and higher tumor grades. Therefore, evaluation of CD44 and CD133 expression profiles as CSCs markers in non-neoplastic liver and HCCs can help in development of novel therapeutic agents for HCC targeting and prevention.

Predictors of histological indication for treatment in HBeAg negative chronic HBV infection.

In this study, we evaluated the relationship of hepatitis B virus (HBV) DNA levels with liver histology and various other liver related parameters and the predictors of histologically active liver disease requiring treatment (Histological activity ≥6 and/or grade ≥2 by Ishak's classification) in patients with HBeAg negative chronic HBV infection. Demographic data, laboratory findings and liver histology findings of patients with no clinical cirrhosis who underwent liver biopsy considering HBeAg negative chronic hepatitis (HBV DNA >2000 IU/mL) were analyzed. Two hundred and fifteen patients were included in this retrospective study. Treatment indication by histologic findings were 85.7%, 61.2%, and 64%, respectively, in group 1 (HBV DNA ≥200 000), group 2 (HBV DNA 20 000-200 000), and group 3 (HBV DNA 2000-20 000 IU/mL) (P = 0.001). Group 1 was different from other groups in terms of aspartate aminotransferase (AST), alanine aminotransferase (ALT), fibrosis stage, necroinflammatory activity, and platelet count. Multiple logistic regression analysis revealed that, advanced age (cut-off was 46 years), higher than normal AST and HBV DNA ≥200 000 IU/mL (compared to group 3) were found to be the predictors of histologically active disease with treatment indication. Conclusively, most of the patients with HBV DNA ≥200 000 IU/mL showed treatment requiring liver injury, but also a significant portion of the patients with HBV DNA 2000-200 000 IU/mL carried an indication for treatment. Although age (>46 years) and AST (>40 IU/L) can be helpful to predict treatment requirement in patients with HBV DNA 2000-200 000 IU/mL, sufficient effort should be made to find out the significant liver damage.

Interferon-related genetic markers of necroinflammatory activity in chronic hepatitis C.

IntroductionChronic hepatitis C (CHC) is a major cause of liver disease worldwide which often leads to progressive liver inflammation, fibrosis, cirrhosis and hepatocellular carcinoma (HCC). CHC displays heterogeneous progression depending on a broad set of factors, some of them intrinsic to each individual such as the patient's genetic profile. This study aims to evaluate the contribution of certain genetic variants of crucial interferon alpha and lambda signaling pathways to the hepatic necroinflammatory activity (NIA) grade of CHC patients.MethodsNIA was evaluated in 119 CHC patients by METAVIR scale and classified as low (NIA = 0–2, n = 80) or high grade (NIA = 3, n = 39). In a candidate gene approach, 64 SNPs located in 30 different genes related to interferon pathways (IL-28B, IFNAR1-2, JAK-STAT and OAS1-3, among others) were genotyped using the Illumina GoldenGate® Genotyping Assay. Statistical association was determined by logistic regression and expressed as OR and 95% CI. Those SNPs significantly associated were further adjusted by other covariates.ResultsSeven SNPs located in IL-28B (rs12979860), JAK1 (rs11576173 and rs1497056), TYK2 (rs280519), OAS1 (rs2057778), SOCS1 (rs33932899) and RNASEL (rs3738579) genes were significantly related to severe NIA grade (p<0.05). Regarding to clinical variables, elevated NIA was notably associated with aspartate aminotransferase (AST) serum levels >40 IU/L (p<0.05) but not with other clinical factors. Multivariate logistic regression analysis of these factors reflected that AST (>40 IU/L), TYK2 rs280519 (G allele) and RNASEL rs3738579 (G allele) were factors independently associated with elevated NIA (p<0.05). AST concentration showed a moderate AUC value (AUC = 0.63), similar to TYK2 (rs280519) and RNASEL (rs3738579) SNPs (AUC = 0.61, both) in the ROC_AUC analysis. Interestingly, the model including all significant variables reached a considerable predictive value (AUC = 0.74).ConclusionThe identified genetic variants in interferon signaling pathways may constitute useful prognostic markers of CHC progression. Further validation in larger cohorts of patients is needed.

Study of histomorphological characteristics and it’s correlation with clinical, biochemical, serological and immunohistochemical parameters in incidentally detected hepatitis B patients

Background India lies in intermediate endemicity zone for hepatitis B virus (HBV) infection and constitutes the second largest global pool of HBV infection worldwide. Hepatitis B has a varied clinical presentation ranging from clinically asymptomatic state to cirrhosis and hepatocellular carcinoma. A significant liver injury can occur, without accompanying elevation in alanine transaminase (ALT) and HBV DNA levels, especially in incidentally detected asymptomatic hepatitis B subjects (IDAHS). Hence, a role of liver biopsy to be incorporated with other investigations is debatable, but important to initiate antiviral therapy. We explored correlation between histomorphological outcomes with various clinical, biochemical, serological and immunohistochemical parameters in IDAHS. Methods Total 113 patients were consecutively selected over a period of 4.5 years. Serological work-up for HBsAg, Anti-HBeAg, Anti-HBeAb, Anti-HBcAb, and HBV DNA levels were done as per resources. A liver biopsy was done in each patient after a written consent. Ishak’s scoring system was used to assess histological parameters. Immunohistochemistry (IHC) was done for HBsAg and HBcAg. Appropriate statistical tests were applied. Results The mean age of the patients was 30 years with a male to female ratio of 3:1. A higher necro-inflammatory activity (NIA >3) correlated with high ALT (>40 U/l), HBV DNA (>10 5 copies/ml) and fibrosis (F ≥2). HBeAg-positive patients had significantly higher NIA and HBV DNA levels. Anti-HBeAb delineated association with ALT (≤40 U/l) and low HBV DNA but more severe fibrosis (F≥2). Steatotic changes were noted in 52.2% biopsies. IHC for HBsAg and HBeAg showed positivity in 82.7% and 39.2% of cases respectively with a significant correlation between membranous pattern of HBsAg staining and serum HBV DNA levels. Conclusion IDAHS represent tip of the iceberg of major HBV infection reservoir. A liver biopsy is a useful additional tool with other parameters to further tailor the therapy in such asymptomatic patients.

Acute presentation of autoimmune hepatitis: a multicentre study with detailed histological evaluation in a large cohort of patients

AimsAlthough liver biopsy is crucial to diagnose and guide treatment decisions, a detailed histological analysis of autoimmune hepatitis (AIH) with clinically acute presentations has not yet been performed. This study...

Interferon-γ-inducible protein-10 in chronic hepatitis C: Correlations with insulin resistance, histological features & sustained virological response.

Background & objectives:One of the multiple factors contributing to virological response in chronic hepatitis C (CHC) is interferon-gamma-inducible protein-10 (IP-10). Its level reflects the status of interferon-stimulated genes, which in turn is associated with virological response to antiviral therapy. The aim of this study was to evaluate the role of serum IP-10 levels on sustained virological response (SVR) and the association of this parameter with insulin resistance (IR) and liver histology.Methods:Two hundred and three consecutive biopsy proven CHC patients were included in the study. Serum levels of IP-10 were determined using ELISA method. IR was evaluated by homeostasis model assessment-IR (HOMA-IR). Histological features were assessed invasively by liver biopsy and noninvasively using FibroTest, ActiTest and SteatoTest. Predictive factors for SVR and their interrelations were assessed.Results:A cut-off value for IP-10 of 392 pg/ml was obtained to discriminate between responders and non-responders. SVR was obtained in 107 patients (52.70%). Area under the receiver operating characteristic curve for SVR was 0.875 with a sensitivity of 91.6 per cent, specificity 74.7 per cent, positive predictive value 80.3 per cent and negative predictive value 88.7 per cent. Higher values of IP-10 were associated with increasing stages of fibrosis (P<0.01) and higher grades of inflammation (P=0.02, P=0.07) assessed morphologically and noninvasively through FibroTest and ActiTest. Significant steatosis and IR were also associated with increased levels of IP-10 (P=0.01 and P=0.02). In multivariate analysis, IP-10 levels and fibrosis stages were independently associated with SVR.Interpretation & conclusions:Our findings showed that the assessment of serum IP-10 level could be a predictive factor for SVR and it was associated with fibrosis, necroinflammatory activity, significant steatosis and IR in patients with chronic HCV infection.